Bispecific antibodies against HER2 and CD3

Abstract

Bispecific antibodies which comprise one antigen-binding region binding to an epitope of human epidermal growth factor receptor 2 (HER2) and one antigen-binding region binding to human CD3, and related antibody-based compositions and molecules, are disclosed. Pharmaceutical compositions comprising the antibodies and methods for preparing and using the antibodies are also disclosed.

The invention claimed is: 
     
       1. A bispecific antibody comprising a first antigen-binding region, a second antigen-binding region, and a first Fc region and a second Fc region, wherein the second antigen-binding region binds an epitope on human CD3 and the first antigen-binding region binds to human epidermal growth factor receptor 2 (HER2), and wherein the first antigen-binding region comprises a heavy chain variable (VH) region and a light chain variable (VL) region selected from the group consisting of:
 (a) a VH region comprising the CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 166, 167 and 168, respectively; and a VL region comprising the CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NO: 170, GAS and SEQ ID NO: 171, respectively; 
 (b) a VH region comprising the CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 173, 174 and 175, respectively; and a VL region comprising the CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NO: 177, DAS, and SEQ ID NO: 178, respectively; 
 (c) a VH region comprising the CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 180, 181 and 182, respectively; and a VL region comprising the CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NO: 184, GAS, and SEQ ID NO: 185, respectively; 
 (d) a VH region comprising the CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 187, 188 and 189, respectively; and a VL region comprising the CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NO: 191, GAS, and SEQ ID NO: 192, respectively; 
 (e) a VH region comprising the CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 194, 195 and 196, respectively; and a VL region comprising the CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NO: 198, GAS, and SEQ ID NO: 199, respectively; and 
 (f) a VH region comprising the CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NOs: 201, 202 and 203, respectively; and a VL region comprising the CDR1, CDR2 and CDR3 sequences set forth in SEQ ID NO: 205, GAS, and SEQ ID NO: 206, respectively, 
 and wherein the first and second Fc regions comprise a sequence independently selected from the group consisting of SEQ ID NOs: 251,254, and 255. 
 
     
     
       2. The bispecific antibody of  claim 1 , wherein the first antigen-binding region comprises VH and VL regions comprising amino acid sequences selected from the group consisting of:
 (a) SEQ ID NOs: 165 and 169, respectively; 
 (b) SEQ ID NOs: 172 and 176, respectively; 
 (c) SEQ ID NOs: 179 and 183, respectively; 
 (d) SEQ ID NOs: 186 and 190, respectively; 
 (e) SEQ ID NOs: 193 and 197, respectively; and 
 (f) SEQ ID NOs: 200 and 204, respectively. 
 
     
     
       3. The bispecific antibody of  claim 1 , wherein the second antigen-binding region comprises VH and VL regions comprising amino acid sequences selected from the group consisting of:
 (a) SEQ ID NOs: 240 and 241, respectively; 
 (b) SEQ ID NOs: 234 and 235, respectively; 
 (c) SEQ ID NOs: 238 and 239, respectively; and 
 (d) SEQ ID NOs: 236 and 237, respectively. 
 
     
     
       4. The bispecific antibody of  claim 1 , which is effector-function deficient. 
     
     
       5. The bispecific antibody of  claim 1 , wherein the first Fc region has an amino acid substitution at a position selected from the group consisting of 409, 366, 368, 370, 399, 405 and 407, and said second Fc region has an amino acid substitution at a position selected from the group consisting of 405, 366, 368, 370, 399, 407, and 409, wherein said first Fc region and said second Fc region are not substituted in the same positions, and wherein the numbering of amino acid positions is according to the EU Index. 
     
     
       6. The bispecific antibody of  claim 5 , wherein said first and second Fc regions, except for the specified mutations, comprise the sequence of SEQ ID NO:247 (IgG1m(a)). 
     
     
       7. The bispecific antibody of  claim 1 , wherein neither said first nor said second Fc region comprises a Cys-Pro-Ser-Cys sequence in the hinge region. 
     
     
       8. The bispecific antibody of  claim 1 , wherein both said first and second Fc regions comprise a Cys-Pro-Pro-Cys sequence in the hinge region. 
     
     
       9. The bispecific antibody of  claim 1 , wherein the first and/or the second Fc-region comprises a mutation removing the acceptor site for Asn-linked glycosylation. 
     
     
       10. A conjugated bispecific antibody of  claim 1 , wherein the antibody is conjugated to a drug, radioisotope, cytokine, or cytotoxic moiety. 
     
     
       11. A pharmaceutical composition comprising the bispecific antibody of  claim 1  and a pharmaceutically acceptable carrier.